Combing Contrast-enhanced Ultrasonography And MMP-9、Endothelin-1Level To Predict The Recurrence Of Hepatocellular Carcinoma After Thermal Ablation

Objective: To investigate the relationship of parameters of contrast enhancedultrasonography (CEUS), levels of MMP-9and Endothelin-1in cancer tissues withrecurrence of HCC after thermal ablation respectively, then analyze the correlationbetween them, in order to find effective characteristic parameters to predict recurrenceof HCC after microwave ablation (MWA) or radiofrequency ablation (RFA) early andnoninvasively.Materials and methods:⑴All the enrolled patients should meet the following criteria: HCC patients confirmedby ultrasound-guided biopsy, single lesion,5cm or less of the maximum diameter,without vascular invasion and extrahepatic metastasis, class A or B of Child-Pugh, noother serious diseases such as heart, brain, kidney and blood vessels. All of the patientsshouldn’t accept other therapies before treatment with MWA or RFA. Patients enrolledshould participate in this study voluntarily and sign informed consent before treatment.⑵The study totally enrolled40HCC patients underwent MWA or RFA from March2006to July2012in our hospital. CEUS were performed on all of them beforetreatment, then quantitative analyses were carried out using Sonoliver software to obtaincharacteristic parameters. Further, statistical analyses were made on those parametersbetween the recurrence group and non-recurrence group to screen recurrence-relatedindexes.⑶HCC tissues of22patients who were selected randomly were obtained throughultrasound-guided biopsy before treatment. The levels of MMP-9and Endothelin-1 proteins were detected by immunohistochemical staining for all of the22specimens.Image Pro-plus5.1software was used to measure the integrated optical density (IOD)of them, and the values were analyzed statistically between the two groups.⑷To analyse the correlation between characteristics parameters of CEUS and thelevels of MMP-9, Endothelin-1, so as to combine them to predict recurrence of HCCafter microwave ablation (MWA) or radiofrequency ablation (RFA).Results:⑴No significant differences were detected in sex, age, HBV/HCV infection, AFPlevels and differentiation degree before treatment between recurrence group andnon-recurrence group.⑵There are significant differences in parameters of tIMAX, IMAXR, tRT, tTTP andWT between the two groups, and all of the values of the recurrence group were smallerthan the non-recurrence one. Multivariate analysis showed that IMAXR was a riskfactor of recurrence (OR=0.111；95%CI，0.0150.838；P=0.033), the smaller of theIMAXR, the higher risk of recurrence.⑶An analysis of the immunohistochemical staining results showed that the expressionlevels of MMP-9in cancer tissues were statistically significant different between thetwo groups, which were higher for the recurrence group. However, no statisticaldifference was detected in the expression levels of Endothelin-1between them. Further,logistic regression analysis showed that none of them were risk factors for recurrence.⑷Correlation analysis between characteristic parameters of CEUS and expressionlevels of MMP-9, Endothelin-1in cancer tissues revealed that tRT and tTTP wereassociated with the expression levels of Endothelin-1moderately [r was0.438and0.448respectively, p＜0.05], while tIMAX, IMAXR and WT showed no correlationwith the levels of Endothelin-1. In addition, there was no correlation between levels ofMMP-9and all the parameters.Conclusions:⑴In all of the parameters of CEUS, IMAXR is a risk factor for recurrence, the smallerof the IMAXR, the higher risk of recurrence. ⑵The expression levels of MMP-9, Endothelin-1in cancer tissue are not risk factorsfor recurrence.⑶The parameters of tRT and tTTP are moderately associated with the expressionlevels of Endothelin-1in cancer tissues, which reveals that some characteristicparameters of CEUS can reflect the expression levels of angiogenesis-related proteins intumor tissues to some extent.⑷Contrast-enhanced ultrasound combining with quantitative analysis can not onlyshow the hemodynamic of the HCC, but also could reflect the changes of expression oftumor proteins on a molecular level. It can be expected to become a powerful tool formonitoring and predicting biological behaviors of HCC, so as to provide guidance forindividualized interventions.